mBMMs (1.0 × 105 cells/well) were harvested from a 12-well plate and treated with RANKL and M-CSF (100 ng/ml each) in the presence and absence of TER (10 μM) for 2 days. Total ribonucleic acid (RNA) was then isolated using the RNeasy® Mini Kit (Qiagen, Hilden, Germany), followed by deoxyribonucleic acid (DNA) removal using an RNase-free DNase Kit (Qiagen). A total of 1 μg of high-quality total RNA was then reverse-transcribed using the SuperScript® III First-Strand Synthesis System (Thermo Fisher Scientific, Waltham, MA, United States). Amplification reactions were performed using SYBR® Green PCR Master Mix (Thermo Fisher Scientific). Up to 1 ng of complementary DNA was then amplified using specific primers. The reactions were performed using a 7,300 Real-Time PCR System (Thermo Fisher Scientific). The ratios of messenger RNA levels to control values were calculated using the ΔCt method (2−ΔΔCt). All data were normalized to the housekeeping control gene, β-actin. The PCR conditions used were as follows: 10 min at 95°C, followed by 40 cycles of 15 s at 95°C, and 60 s at 60°C. The primers used were as follows: 5′-TGG​GCC​TCC​ATA​TGA​CCT​CGA​GTA​G-3′ (forward) and 5′-TCA​AAG​GCT​TGT​AAA​TTG​GAG​GAG​T-3′ (reverse) for osteoclast stimulatory transmembrane protein (Ocstamp), 5′-CTA​GCT​GGC​TGG​ACT​TCA​TCC-3′ (forward) and 5′-TCA​TGC​TGT​CTA​GGA​GAC​CTC-3′ (reverse) for dendritic cell–specific transmembrane protein (Dcstamp), 5′-TGC​AGA​CAA​CTC​TTG​GTT​GG-3′ (forward) and 5′-TCG​GTT​TCT​TCT​CCT​CTG​GA-3′ (reverse) for calcitonin receptor (Calcr), 5′-TCA​GAT​CTC​TTC​AAG​GCT​GTG​CTG-3′ (forward) and 5′-GTG​CCA​AAT​GAG​TTC​AGA​GTG​ATG-3′ (reverse) for v-type protein ATPase subunit d2 (Atp6v0d2), 5′-CTG​CTG​GTA​ACG​GAT​CAG​CTC​CCC​AGA-3′ (forward) and 5′-CCA​AGG​AGC​CAG​AAC​CTT​CGA​AAC​T-3′ (reverse) for osteoclast-associated receptor (Oscar), 5′-ATG​CCA​GCG​ACA​AGA​GGT​TC-3′ (forward) and 5′-TGG​TTT​CCA​GCC​AGC​ACA​TAC-3′ (reverse) for Trap (Acp5), 5′- TGA​CCA​CTG​CCT​TCC​AAT​ACG-3′ (forward) and 5′-TGC​ATT​TAG​CTG​CCT​TTG​CC-3′ (reverse) for cathepsin-K, 5′-TGT​GTG​CCT​GGT​GCT​CAG​A-3′ (forward) and 5′-AGC​AGG​TTC​TCC​TTC​AGG​TTA​CA-3′ (reverse) for integrin β3 (Itgb3), 5′-CCA​GTC​AAG​AGC​ATC​AGC​AA-3′ (forward) and 5′-AAG​TAG​TGC​AGC​CCG​GAG​TA-3′ (reverse) for Proto-oncogene c-fos (c-fos), and 5′-TAG​CGG​AAC​CGC​TCA​TTG​CC-3′ (forward) and 5′-TTC​ACC​CAC​ACT​GTG​CCC-3′ (reverse) for β-actin (Kwak et al., 2010; Choi et al., 2012; Lee et al., 2012; Ma et al., 2014; Shimada-Sugawara et al., 2015; Nakagawa et al., 2020).

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